KR20200089523A - Method for increasing growth and metabolite contents of pirodela polyrhiza (L.) Schleid - Google Patents

Abstract

The present invention relates to a method for increasing the growth and metabolite content of Spirodela polyrhiza (L.) Schleid, comprising: 1) a step of placing seedlings of Spirodela polyrhiza (L.) Schleid in a nutrient solution; and 2) a step of culturing the seedlings of Spirodela polyrhiza (L.) Schleid placed in the nutrient solution of step 1) under light-emitting diode (LED) light conditions. Since the method increases the fresh weight, the dry weight, and the number of fronds of Spirodela polyrhiza (L.) Schleid and increases the content of metabolites, the method can be usefully used for mass proliferation of Spirodela polyrhiza (L.) Schleid and production of Spirodela polyrhiza (L.) Schleid with enhanced metabolite content.

Description

Method for increasing growth and metabolite contents of pirodela polyrhiza (L.) Schleid}

The present invention is duckweed ( Spirodela polyrhiza (L.) Schleid ) is related to a method for enhancing the growth and metabolite content.

Duckweed[ Spirodela polyrhiza (L.) Schleid ] is a small aquatic plant that breeds antelope (Lemnaceae) and has a higher growth rate than other plants (Vermaat, JE, & Hanif, KM (1998), Performance of common duckweed species ( Lemnaceae ) and the waterfern Azolla filiculoides on different types of waste water. Water Research, 32, 2569-2576). By using these advantages of duckweed, it is also used for water quality improvement such as sewage treatment system and improvement of eutrophication (Dalu, JM, & Ndamba, J. (2003). Duckweed based wastewater stabilization ponds for wastewater treatment (a low cost technology for small urban areas in Zimbabwe).Physics and Chemistry of the Earth, Parts A/B/C, 28(20), 1147-1160), contain relatively low fiber and high protein and are also used as feed for livestock (Landolt E (1986), The family of Lemnaceae: A monographic study.Vol. 1.Biosystematic investigations in the family of duckweeds (Lemnaceae) (vol. 2).VeroE.Geobot.Inst.ETH Stift.Ruebel 71, 1-566; Oron G. et al. (1986), J. Environ. Eng. (Am. Soc. Civil Eng.) 112, 247-263; Whitehead AJ et al. (1987), In Aquatic Plants for Water Treatment and Resource Recovery, eds.KR Reddy and WH Smith, pp. 697-703.Magnolia Publ. Orlando, FL, USA).

According to research by Culley DD and Epps EA (J. Wat. Poll. Control Fed. 45, 337-347, 1973), duckweed grown in a specific sewage has a protein content of 40% based on dry weight. And 30-50 mg/lb of carotene, indicating the efficacy of vitamin A, was detected.

As described above, although the value of utilization of duckweed is increasing, studies on the mass growth of duckweed and the content of secondary metabolites have been insufficient. Therefore, it is necessary to develop a method of promoting the growth of duckweed and enhancing the content of metabolites.

The purpose of the present invention is duckweed ( Spirodela polyrhiza (L.) Schleid ) to provide a method for enhancing the growth and content of metabolites.

In order to achieve the above object, the present invention is 1) duckweed ( Spirodela polyrhiza (L.) Schleid ) to the seedling of the seedling; And 2) culturing the seedlings of duckweed dentured in the nutrient solution of step 1) under light emitting diode (LED) light conditions.

When seedlings of duckweed in the present invention are dentified in a nutrient solution containing 20% of nitrogen, phosphoric acid and potassium, respectively, and cultured by adjusting the light-emitting diode (LED) red light, green light, blue light or white light ratio. Since it was confirmed that the number of live weight, building weight, and fronds of the duckweed increased and the content of metabolites increased, the method can be usefully used to produce duckweed with a large amount of duckweed and an enhanced metabolite content.

1 is a view showing the appearance of duckweed cultured according to the concentration conditions of the nutrient solution and the light-emitting diode red light, green light or blue light ratio conditions.
(R: red light (660 nm); G: green light (525 nm); B: blue light (450 nm); Solar: sunlight; Tap: tap water (control).
FIG. 2 is a diagram showing the results of fresh weight and dry weight of duckweed cultured according to the concentration conditions of the nutrient solution and the light ratio of the light emitting diode red light, green light or blue light.
-Fw-con: weight of control group; Fw-500: Live weight of nutrient solution 500 ppm treated group; Fw-1000: Live weight of the nutrient solution 1,000 ppm treatment group; Fw-2000: Live weight of nutrient solution 2,000 ppm treated group;
-Dw-con: In the control building; Dw-500: nutrient solution 500 ppm of treatment group among buildings; Dw-1000: Among the buildings of the nutrient solution 1000 ppm treatment group; Dw-2000: Among the buildings of the nutrient solution 2000 ppm treatment group;
-con: sunlight; R8G2: luminous intensity ratio of red light:green light=8:2; R6B4: red light: blue light ratio = 6:4; R8B2: red light: blue light ratio = 8:2; R6G2B2: red light: green light: blue light intensity ratio = 6:2:2;
-a, b, c and d: Duncan's multiple test results using SAS statistics program.
3 is a view showing the results of the number of fronds of duckweed cultured according to the concentration conditions of the nutrient solution and the light-emitting diode red light, green light or blue light ratio conditions.
-control: sunlight; R8G2: luminous intensity ratio of red light:green light=8:2; R6B4: red light: blue light ratio = 6:4; R8B2: red light: blue light ratio = 8:2; R6G2B2: red light: green light: blue light intensity ratio = 6:2:2;
-a, b, c and d: Duncan's multiple test results using SAS statistics program.
4 is a diagram showing the results of the number of fronds of duckweed cultured according to the concentration conditions of the nutrient solution and the luminous diode red light, green light or blue light ratio conditions.
-control: sunlight; R8G2: luminous intensity ratio of red light:green light=8:2; R6B4: red light: blue light ratio = 6:4; R8B2: red light: blue light ratio = 8:2; R6G2B2: red light: green light: blue light intensity ratio = 6:2:2;
-a, b, c and d: Duncan's multiple test results using SAS statistics program.
5 is a diagram showing the results of live weight of duckweed cultured under conditions of nutrient solution concentration of 1,000 ppm and light emitting diode red light, green light and blue light ratio.
-R8G4B5: red light: green light: blue light ratio ratio = 8:4:5; R7G4B6: ratio of luminous intensity of red light: green light: blue light = 7:4:6; R6G4B7: red light: green light: blue light ratio ratio = 6:4:7; R5G4B8: red light:green light:blue light ratio =5:4:8).
FIG. 6 is a diagram showing the metabolite content of duckweed cultured according to conditions of nutrient solution concentration of 1,000 ppm and light emitting diodes of red light, green light and blue light.
-Per weight: the content of metabolites in the same weight; X Fresh weight: the total metabolites content in the body of total duckweed produced;
-R8G4B5: red light: green light: blue light ratio ratio = 8:4:5; R7G4B6: ratio of luminous intensity of red light: green light: blue light = 7:4:6; R6G4B7: red light: green light: blue light ratio ratio = 6:4:7; R5G4B8: red light:green light:blue light ratio =5:4:8).
Figure 7 is a diagram showing the results of living and building of duckweed cultured under conditions of nutrient solution concentration of 1,000 ppm and light emitting diode red light, green light, blue light or white light.
-W9R1: luminous intensity ratio of white light:red light=9:1; White: White light; W3R1: White light:Red light intensity ratio = 3:1; R8G4B5: red light:green light:blue light ratio =8:4:5; R6B4: Red light: blue light ratio = 6:4).
-a, b and c: Duncan's multiple test results using SAS statistics program.
FIG. 8 is a diagram showing a CAD (Charged Aerosol Detector) chromatogram analyzing the components of duckweed cultured according to conditions of nutrient solution concentration of 1,000 ppm and light emitting diode red light, green light, blue light or white light.
-Standard: Analysis results of duckweed grown outdoors; Peak 1: Luteolin 8-C-glucoside; Peak 2: Apigenin 6-C-glucoside; Peak 3: Luteolin 7-O-glucoside; Peak 4: Apigenin 7-O-glucoside; Peak 5: Luteolin 8-C-(2''-O-feruloyl)-glucoside; Luteolin 8-C-(2''-O-feruloyl)-glucoside; Peak 6: Apigenin 8-C-(2''-O-feruloyl)-glucoside (Apigenin 8-C-(2''-O-feruloyl)-glucoside); Peak 7: Luteolin.
9 is a diagram showing a UV chromatogram analyzing the components of duckweed cultured according to conditions of a nutrient solution concentration of 1,000 ppm and light-emitting diode red light, green light, blue light or white light.
10 is an UPLC-PDA-QTof-MS (ultra performance liquid chromatography-photodiode detector-quadrupole) analyzing the composition of duckweed cultured according to conditions of nutrient solution concentration 1,000 ppm and light emitting diode red light, green light, blue light or white light. /time of flight-mass spectrometry).
11 is a diagram showing the metabolite content of duckweed cultured under conditions of nutrient solution concentration of 1,000 ppm and light-emitting diode red light, green light, blue light or white light.
-W9R1: luminous intensity ratio of white light:red light=9:1; White: White light; W3R1: White light:Red light intensity ratio = 3:1; R8G4B5: red light:green light:blue light ratio =8:4:5; R6B4: Red light: blue light ratio = 6:4).
-Per weight: the content of metabolites contained in the same weight (100 mg); X Dry weight: Total metabolite content in the building of the total duckweed produced.

Hereinafter, the present invention will be described in detail.

The present invention is 1) duckweed ( Spirodela seeding the seedlings of polyrhiza (L.) Schleid ) in nutrient solution; And 2) culturing the seedlings of the duckweed dentured in the nutrient solution of step 1) under light-emitting diode (LED) light conditions to provide a method for enhancing the growth and metabolite content of duckweed.

The nutrient solution of step 1) may contain nitrogen, phosphoric acid and potassium at 10 to 30%, respectively. Specifically, the nutrient solution of step 1) may include nitrogen, phosphoric acid and potassium in 10 to 25%, 15 to 30%, 15 to 25%, or 17 to 23%, respectively. According to an embodiment of the present invention, the nutrient solution may include nitrogen, phosphoric acid, and potassium in 20%, respectively.

As described above, the concentration of the nutrient solution containing nitrogen, phosphoric acid and potassium may be 800 to 2,200 ppm. Specifically, the concentration of the nutrient solution may be 800 to 2,000 ppm, 800 to 1,800 ppm, 800 to 1,600 ppm, 800 to 1,400 ppm, or 800 to 1,200 ppm. According to an embodiment of the present invention, the concentration of the nutrient solution may be 1,000 ppm.

The light condition of the light emitting diode of step 2) may be a mixture treatment of two or more selected from the group consisting of red light, green light, blue light and white light.

Specifically, the light condition of the light emitting diode of the step 2) may be a red light: blue light ratio of 5 to 9: 1 to 5, and more specifically, a red light: blue light ratio of 6 to 9: 1 to 5, 6 It may be 9 to 1 to 4, 7 to 9: 1 to 5, 7 to 9: 1 to 4, or 7 to 9: 1 to 3. According to an embodiment of the present invention, the light condition of the light emitting diode of step 2) may be 8:2 with a red light:blue light ratio.

In addition, the light condition of the light emitting diode of the step 2) may be a red light: green light: blue light ratio of 4 to 9: 3 to 5: 4 to 9, and more specifically, the red light: green light: blue light has a light ratio of 5 to 9: 3 to 5: 4 to 9, 5 to 9: 3 to 5: 4 to 8, 5 to 9: 3 to 5: 4 to 7, 5 to 9: 3 to 5: 4 to 6, 6 to 9: It may be 3 to 5: 4 to 7, 6 to 9: 3 to 5: 4 to 6, 7 to 9: 3 to 5: 4 to 7, or 7 to 9: 3 to 5: 4 to 6. According to an embodiment of the present invention, the light condition of the light emitting diode of step 2) may be 8:4:5 in the ratio of red light: green light: blue light.

In addition, the light condition of the light emitting diode of the step 2) may be a white light: red light ratio of 2 to 10: 1, more specifically white light: red light ratio of 3 to 10: 1, 4 to 10: 1, It may be 5 to 10:1, 6 to 10:1, 7 to 10:1, or 8 to 10:1. According to an embodiment of the present invention, the light condition of the light emitting diode of step 2) may be in a ratio of white light:red light of 9:1.

The red light may have a wavelength of 640 to 680 nm, 650 to 680 nm, 655 to 680 nm, 640 to 670 nm, 640 to 665 nm, 650 to 670 nm, or 655 to 665 nm.

The green light may have a wavelength of 505 to 545 nm, 515 to 545 nm, 520 to 545 nm, 505 to 535 nm, 505 to 530 nm, 515 to 535 nm, or 520 to 530 nm.

The blue light may have a wavelength of 430 to 470 nm, 440 to 470 nm, 445 to 470 nm, 430 to 460 nm, 430 to 455 nm, 440 to 460 nm, or 445 to 455 nm.

In addition, the light condition of the light emitting diode of step 1) may be 600 to 700 mol·m ^-2 ·s ^-1 . Specifically, the light intensity is 620 to 700 mol·m ^-2 · s ^-1 , 640 to 700 mol·m ^-2 · s ^-1 , 650 to 700 mol·m ^-2 · s ^-1 , 650 to 690 mol·m ^{It may be -2} ·s ^-1 , or 660 to 680 mol·m ^-2 ·s ^-1 .

The culture of step 2) may be performed in a closed-type plant production system.

In addition, the culture of step 2) may be made at a temperature of 22 to 32° C., a relative humidity of 30 to 70% and a carbon dioxide (CO ₂ ) concentration of 700 to 1,500 ppm.

In addition, the culture of step 2) may be performed for 7 to 15 days, specifically 8 to 15 days, 9 to 15 days, 7 to 14 days, 7 to 13 days, 8 to 14 days, or 9 to 13 days It can be one day.

In a specific embodiment of the present invention, the present inventors have a concentration condition (0, 500, 1000 or 2000 ppm) of nutrient solution and a light emitting diode red light, green light or blue light ratio condition (red light: green light = 8:2; red light: blue light = 6:4; red light:blue light=8:2; red light:green light:blue light=6:2:2) As a result of cultivating duckweed, the fresh weight and dry weight of the duckweed are nutrient concentration. It was confirmed that the concentration increased at 1,000 and 2,000 ppm (see FIG. 2 ), and the number of fronds increased most at 1,000 ppm (see FIGS. 1 and 3 ). In addition, the body weight of duckweed showed excellent tendency regardless of the concentration of nutrient solution when treated with 8:2 ratio of red light and blue light, and in the building, when treated with 8:2 ratio of red light and blue light, the concentration of 1,000 ppm of nutrient solution was observed. It was confirmed to be the highest (see FIG. 2). The number of fronds appears to increase when treated with 8:2 and 6:4 ratios of red and blue light (see FIGS. 1 and 4), and the growth of duckweed is enhanced as the ratio of red light in the light emitting diode increases, and an appropriate nutrient solution The concentration of was confirmed to be 1,000 ppm.

In addition, the present inventors have a condition of nutrient solution concentration of 1,000 ppm and light emitting diode red light, green light and blue light ratio conditions (red light: green light: blue light = 8:4:5; red light: green light: blue light = 7:4:6; red light: Green light: blue light = 6:4:7; red light: green light: blue light = 5:4:8) differently, the result of cultivation of duckweed, the red light: green light: blue light ratio is 8:4:5 The most increased (see Fig. 5) and the content of the metabolites increases the most (see Fig. 6), so the growth and metabolites of duckweed in conditions of red light:green light:blue light and blue light at 1,000 ppm concentration and 8:4:5 ratio It was confirmed that the content was effectively enhanced.

In addition, the present inventors nutrient solution concentration conditions of 1,000 ppm and the light emitting diode red light, green light, blue light or white light luminous ratio conditions (white light: red light = 9: 1; white light; white light: red light = 3: 1; red light: green light: blue light = 8:4:5; red light:blue light=6:4)), and as a result of cultivation of duckweed, when combined with white light and red light, the body weight and building weight increased (see Fig. 7) and the content of metabolites increased. Therefore, (see FIG. 11), it was confirmed that the growth and metabolite content of the duckweed are effectively enhanced in the condition of 1,000 ppm nutrient solution and 3 to 9:1 ratio of white light: red light.

Therefore, the above method can be usefully used to produce duckweed with a large amount of metabolites and a large amount of metabolites, since it increases the number of metabolites and increases the number of live weights, building weights, and fronds.

Hereinafter, the present invention will be described in detail by examples.

However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited by the following examples.

< Example 1> Culture of duckweed

After adding tap water to a height of about 5 cm in 9 square boxes (18 cm x 23 cm x 9 cm), the rice balls (Spirodela polyrhiza (L.) Schleid .) Was transplanted, and after 19 days, individuals with similar frond size and three fronds were selected and used as materials for the following experiment.

< Example 2> Nutrient solution Concentration and Mineral Cultivation of duckweed with different conditions

In order to confirm the effect of promoting the growth of duckweed according to the nutrient concentration and light quality conditions, the frog rice was cultured by varying the treatment concentration of the water-soluble fertilizer and the light treatment condition.

First, water, water, and fertilizers containing 20% of nitrogen, phosphoric acid, and potassium, respectively (Multifeed 20-20-20 with micronutrient, Haifa Chemical, Haifa, Israel) were prepared at 500, 1000, and 2000 ppm concentrations, and used as a nutrient solution. Each prepared nutrient solution was put into a petri dish 100 mL each, and 10 pieces of duckweed selected in Example 1 were dentured, and this was sealed plant production system (temperature 27.4±2.2°C, relative humidity 51.6±16.1%, CO ₂ 1158.9±336.2 ppm) and cultured under a light emitting diode (LED). Light-emitting diode conditions are a total of four conditions, using a light-emitting diode light source of red light (660 nm), blue light (450 nm), and green light (525 nm), the ratio of red light: green light (R8G2), 6: at a ratio of 8:2 light intensity. Red light:blue light (R6B4) at a ratio of 4 light intensity, red light:blue light (R8B2) at a ratio of 8:2 light intensity, and red light:green light:blue light at a 6:2:2 light intensity ratio were created. The luminous intensity was uniformly adjusted to 669.6±2 μmol·m ^-2 ·s ^-1 on the surface of the frond of the duckweed using the LED quantum sensor (MQ-500, Apogee Inst. Logan, UT, USA). It was set to /8h (light/dark). As a control group, a petri dish containing tap water (0 ppm of nutrient solution) was used, and solar light was placed in a glass greenhouse (temperature 31.2±4.2° C., relative humidity 59.0±14%, CO ₂ 448.5±48.7 ppm).

In order to measure the conditions inside the closed plant production system, temperature, humidity, and CO ₂ sensors (SH-VT250, SOHA Tech, Seoul, Korea) and CO ₂ sensors (GMP222, Vaisala Co., Helsinki, Finland) are used as data loggers ( CR1000, Campbell Scientific, Logan, UT, USA) to continuously record the internal conditions, and to measure the internal conditions of the glass greenhouse, a temperature, humidity, and CO ₂ sensor (SH-VT250, SOHA Tech) was used. .

< Experimental Example 1> Effective for promoting the growth of duckweed Nutrient solution Check density and light conditions

According to Example 2, the growth rate of duckweed with different nutrient concentration and light quality was evaluated. The degree of growth was evaluated using fresh weight, dry weight, and number of fronds of duckweed.

<1-1> Live weight (fresh weight) and Building (dry weight)

The weight of the duckweed refers to the weight of the duckweed that has not been dried, and the weight of the duckweed obtained by drying at a temperature of 15℃ and humidity of 10%. The live weight and building weight of the duckweed cultured under each condition were measured.

As a result, in the experimental group treated with nutrient solution, both the live weight and the building weight increased than the control group cultured under the tap water condition, and the live weight and the building weight of the duckweed cultured in the nutrient solution under 1,000 ppm or 2,000 ppm conditions were higher than in the 500 ppm condition (Fig. 2). In addition, the experimental group treated with the light-emitting diode also increased in both live weight and building weight than the control group under the sunlight condition. In the case of living weight, the ratio of red light:blue light in the ratio of 8:2 was excellent regardless of the concentration of nutrient solution, and in the case of building, it was the highest when the concentration of nutrient solution was 1,000 ppm in the condition of red light:blue light in the ratio of 8:2. There was no significant difference between the experimental groups under other conditions.

<1-2> Number of fronds

The number of fronds of duckweed cultured in each condition was measured.

As a result of confirming the number of foliar bodies according to the concentration of nutrient solution, the experimental group treated with nutrient solution showed that the number of foliar bodies of duckweed increased than that of the control group cultured under tap water conditions. (Figures 1 and 3). In addition, as a result of confirming the number of fronds of duckweed according to the light quality conditions, the experimental group treated with light-emitting diodes increased the number of fronds than the control of the sunlight condition, and the ratio of red light:blue light in the ratio of 8:2 and 6:4 compared to other light conditions. It was shown to be effective in increasing the number of fronds (FIGS. 1 and 4 ).

Through the above results, it was confirmed that the growth of duckweed is enhanced as the proportion of red light in the light emitting diode increases, and the proper nutrient concentration is 1,000 ppm.

< Example 3> Red light , Blue light And Green Cultivation of different proportions of duckweed

The purpose of this study was to confirm the effect of promoting the growth of duckweed according to the ratio of the luminous intensity of the red light, blue light, and green light.

Three duckweed seedlings (0.0125 g) are placed in a petri dish containing 1,000 ppm of nutrient solution, and the ratio of luminosity of red light, green light, and blue light is 8:4:5, 7:4:6, 6:4:7, and 5, respectively. The frog rice was cultured in the same manner as in Example 2, except that the frog rice was cultured for 12 days under the light condition of :4:8.

< Experimental Example 2> Photonic conditions effective for promoting the growth of duckweed ( Red light , Blue light And Green light ) Confirm

According to Example 3, the growth ratio of the cultivated duckweed was evaluated by varying the luminous intensity ratios of three types of red light, blue light, and green light. The degree of growth was evaluated by the fresh weight of duckweed and the content of metabolite of duckweed.

<2-1> Live weight (fresh weight)

The live weight of duckweed was measured on days 0, 4, 6, 8, 10 and 12 of culture.

As a result, in the same tendency as in the result of Experimental Example 1, in the red light:green light:blue light condition of 8:4:5 where the ratio of red light is high, the body weight of duckweed was the highest under the conditions of 12 days of culture, and the weight of the live weight was about 15.4 g on the 12th day of culture. On day 0 of culture, the weight of duckweed was increased by about 1232 times (0.0125 g) (Fig. 5).

<2-2> Metabolite content

To measure the metabolites of duckweed, an extract of duckweed was prepared and the content of metabolites was measured using ultra performance liquid chromatography (UPLC).

Specifically, each cultured duckweed was shaded to remove moisture, and 4 mL of 80% methanol was added to 100 mg of dry weight of the duckweed and repeatedly extracted three times at room temperature using an ultrasonic crusher for 15 minutes. The extract was filtered once with a 0.25 mm membrane filter and concentrated under reduced pressure to obtain duckweed methanol extract (15.3 to 18.6 mg). The components of the obtained methanol extract were analyzed by ultra performance liquid chromatography (UPLC). Specifically, the duckweed extract was filtered once with a 0.25 mm membrane filter for UPLC and then filtered after mounting a column (Waters BEH C18 column, 2.1 × 100 mm, 1.7 μm) on a UPLC device (Waters UPLC-QTOF-MS). Each fraction was loaded in an amount of 2 μl. At this time, acetonitrile + 0.1% formic acid/water + 0.1% formic acid was used as the solvent, and the elution rate was 0.4 ml/min. The gradient of the mobile phase was performed as shown in Table 1 below. At this time, using a UV, CAD (Charged Aerosol Detector), MS (Mass Spectrometry) as a detector, the metabolites separated from the UPLC were chromatographed to confirm the degree of separation of each metabolite, and the content of each metabolite was calculated.

As a result, there was no significant difference in the per weight of metabolites contained in the same weight of duckweed, but the content of the total metabolites (x fresh weight) in the body of the total amount of produced duckweed It was found that the ratio of red light:green light:blue light with a high ratio of red light was the highest in the condition of 8:4:5 (Fig. 6).

Through the above results, when seedlings of duckweed were dentured at a concentration of 1,000 ppm of nutrient solution and treated with red light:blue light:green light in a ratio of 8:4:5, it was confirmed that growth and metabolites were most prominent when cultivated duckweed.

Time(min) Elution rate (mL/min) %A %B 0.00 0.400 95 5 0.73 0.400 95 5 10.11 0.400 50 50 13.32 0.400 0 100 12.19 0.400 0 100 12.40 0.400 95 5 14.28 0.400 95 5

(A: acetonitrile + 0.1% formic acid; B: water + 0.1% formic acid)

< Example 4> White light and Red Cultivation of different proportions of duckweed

In order to find an economic cultivation method for duck rice, we tried to confirm whether the commercial white light-emitting diode is used as the primary light and the growth effect of the frog rice when red light is used as the auxiliary light.

Specifically, frog rice was cultured for 10 days in the same manner as in Example 3-1, except that the frog rice was cultured under light conditions in which the luminosity ratios of white light and red light were 9:1 and 3:1, respectively. As a control, the light conditions that were effective in increasing the production of duckweed in Experimental Examples 1 and 2 were 8:4:5 red light: green light: blue light condition (R8G4B5), 6:4 red light: blue light condition (R6B4) and white light alone conditions. Duckweed was used under (White).

< Experimental Example 3> Effective light conditions (white light and Red light ) Confirm

According to Example 4, the growth rate of the duckweed cultured by varying the ratio of white light and red light was evaluated. The degree of growth was evaluated for fresh weight, dry weight, and metabolite content of duckweed.

<3-1> Live weight (fresh weight) and Building (dry weight)

The live weight and building weight of the duckweed cultured under each condition were measured.

As a result of measuring the live weight, when the white light and the red light were treated at 9:1, the increase in the live weight was 672%, and when the white light and the red light were treated at 3:1, the increase in the live weight was found to be 612%. It was confirmed that the combination of white light and red light was more effective in promoting the growth of duckweed than the condition of blue light = 8:4:5 and white light alone. Even in the measurement result among buildings, the weight of the building (2.06 g) was highest when white light and red light were treated at 9:1 (Fig. 7).

<3-2> Metabolite content

In the same manner as in Experimental Example 2-2, the metabolite content of each duckweed cultured in Example 4 was measured.

Specifically, the separation degree of metabolites was confirmed by using UPLC (FIGS. 8 to 10), and the components corresponding to the peaks confirmed by chromatography are shown in [Table 2]. The metabolite content of duckweed cultivated in a closed system using light-emitting diodes was compared, and the relative content (area %) for metabolites according to the light-emitting diode ratio was analyzed.

As a result, when the ratio of white light:red light was 3 to 9:1, the proliferation rate of duckweed was the highest, and the metabolites of duckweed cultured under different light-emitting diode conditions were metabolites of duckweed (standard product) grown in the natural state. It was confirmed that the same major metabolite as the product was produced (Table 3). Based on the results in Table 3, as a result of analyzing the relative content of metabolites according to the light-emitting diode conditions, the content (per weight) of metabolites contained in duckweed of the same weight is about 18.5% in white light:red light condition of 3:1 The highest, the red light:green light:blue light=8:4:5 confirmed in Experimental Example 2 had the lowest content of metabolites, and the total amount of metabolites contained in the building of the total duckweed produced (x dry weight) ) Was highest in the 9:1 white light:red light condition (Fig. 11).

Through the above results, when the seedlings of duckweed were dentured at a concentration of 1,000 ppm of nutrient solution and treated with white light:red light in a ratio of 3:1 to 9:1, it was confirmed that the growth and metabolites were most prominent when cultivated duckweed.

Peak [MH] ^- UV (nm) MS/MS Molecular formula Compound name One 447.0947 210, 349 327, 297 C ₂₁ H ₂₀ O ₁₁ Luteolin 8-C-glucoside
(Luteolin 8-C-glucoside) 2 431.1017 215, 337 311, 283, 117 C ₂₁ H ₂₀ O ₁₀ Apigenin 6-C-glucoside
(Apigenin 6-C-glucoside) 3 447.0969 206, 347 285, 256 C ₂₁ H ₂₀ O ₁₁ Luteolin 7-O-glucoside
(Luteolin 7-O-glucoside) 4 431.1021 208, 338 268, 240 C ₂₁ H ₂₀ O ₁₀ Apigenin 7-O-glucoside
(Apigenin 7-O-glucoside) 5 623.1391 216, 334 447, 429, 327, 309, 297 C ₃₁ H ₂₈ O ₁₄ Luteolin 8-C-(2''-O-feruloyl)-glucoside
(Luteolin 8-C-(2''-O-feruloyl)-glucoside) 6 607.1483 216, 330 413, 311, 293, 283 C ₃₁ H ₂₈ O ₁₃ Apigenin 8-C-(2''-O-feruloyl)-glucoside
(Apigenin 8-C-(2''-O-feruloyl)-glucoside) 7 285.0430 207, 348 151, 133 C ₁₅ H ₁₀ O ₆ Luteolin

sample Proliferation rate
(ship) ratio peak 1 peak 2 peak 3 peak 4 peak 5 peak 6 peak 7 W9R1 6.73 1.30 1264959 628119 445281 352172 55689 10219 43266 White 5.16 1.00 726362 468712 779204 320944 35476 9326 38993 W3R1 6.12 1.19 1030811 555898 1224933 395329 49352 5364 46806 R8G4B5 5.67 1.10 1161630 563693 1332041 343156 48733 8967 42816 R6B4 3.80 0.75 777097 402522 870686 256779 32931 4608 26484

Claims (9)

1) Spirodela polyrhiza (L.) Schleid ) to the seedling of the seedling; And
2) A method for enhancing the growth and content of metabolites of duckweed comprising culturing seedlings of duckweed dentured in the nutrient solution of step 1) under light emitting diode (LED) light conditions.
According to claim 1,
The nutrient solution of step 1) is nitrogen, phosphoric acid, and potassium, respectively, containing 10 to 30% of the method for enhancing the growth and content of metabolites of duckweed.
According to claim 2,
The concentration of the nutrient solution is 800 to 2,200 ppm, a method for enhancing the growth and content of metabolites of duckweed.
According to claim 1,
The light-emitting diode light condition of the step 2) is a method of enhancing the growth and content of metabolites of duckweed, wherein the luminous intensity ratio of red light:blue light is 5 to 9:1 to 5.
According to claim 1,
The light-emitting diode light condition of the step 2) is a method of enhancing the growth and content of metabolites of duckweed, wherein the light intensity ratio of red light:green light:blue light is 4-9:3-5:4-9.
According to claim 1,
The light-emitting diode light condition of the step 2) is a method of enhancing the growth and content of metabolites of duckweed, wherein the light intensity ratio of white light:red light is 2 to 10:1.
According to claim 1,
The culture of step 2) is performed in a closed-type plant production system, a method of promoting the growth and content of metabolites of duckweed.
According to claim 1,
The culture of step 2) is carried out at a temperature of 22 to 32° C., a relative humidity of 30 to 70% and a carbon dioxide (CO ₂ ) concentration of 700 to 1,500 ppm. .
According to claim 1,
The culture of step 2) is performed for 7 to 15 days, a method for enhancing the growth and content of metabolites of duckweed.

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